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Issue 4, 2011
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A microfluidic system with optical laser tweezers to study mechanotransduction and focal adhesion recruitment

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Abstract

We present a new method to locally apply mechanical tensile and compressive force on single cells based on integration of a microfluidic device with an optical laser tweezers. This system can locate a single cell within customized wells exposing a square-like membrane segment to a functionalized bead. Beads are coated with extracellular matrix (ECM) proteins of interest (e.g. fibronectin) to activate specific membrane receptors (e.g. integrins). The functionalized beads are trapped and manipulated by optical tweezers to apply mechanical load on the ECM-integrin-cytoskeleton linkage. Activation of the receptor is visualized by accumulation of expressed fluorescent proteins. This platform facilitates isolation of single cells and excitation by tensile/compressive forces applied directly to the focal adhesionvia specific membrane receptors. Protein assembly or recruitment in a focal adhesion can then be monitored and identified using fluorescent imaging. This platform is used to study the recruitment of vinculin upon the application of external tensile force to single endothelial cells. Vinculin appears to be recruited above the forced bead as an elliptical cloud, centered 2.1 ± 0.5 μm from the 2 μm bead center. The mechanical stiffness of the membrane patch inferred from this measurement is 42.9 ± 6.4 pN μm−1 for a 5 μm × 5 μm membrane segment. This method provides a foundation for further studies of mechanotransduction and tensile stiffness of single cells.

Graphical abstract: A microfluidic system with optical laser tweezers to study mechanotransduction and focal adhesion recruitment

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Publication details

The article was received on 07 Oct 2010, accepted on 17 Nov 2010 and first published on 08 Dec 2010


Article type: Paper
DOI: 10.1039/C0LC00487A
Citation: Lab Chip, 2011,11, 684-694
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    A microfluidic system with optical laser tweezers to study mechanotransduction and focal adhesion recruitment

    P. Honarmandi, H. Lee, M. J. Lang and R. D. Kamm, Lab Chip, 2011, 11, 684
    DOI: 10.1039/C0LC00487A

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